AIM:Antioxidants act mainly through two routes:induction of antioxidant enzymes(enzymatic)or direct reaction with reactive oxygen species(ROS)(non-enzymatic),but the one taken by the extract of Ginkgo biloba(EGb)remains to be investigated.In present study,EGb was tested for both of the antioxidant routes in vitro.METHODS:The induction of EGb on two antioxidant enzymes,glutamate cysteine ligase catalytic subunit (GCLC)and glutathione-S-transferage subunit-P1(GST-P1),in cell lines wag detected by Western-blot.The effects of EGb on superoxide anion radical(O2·-),hydroxyl radical(OH·),rat erythrocyte hemolysis and lipid peroxidation of rat liver homogenate were determined by activity methods respectively.RESULTS:EGb was dem onstrated significantly to induce the two antioxidant enzymes(GCLC and GST-P1),directly scavenge superoxide anion radical(O2·-),hydroxyl radical(OH·)and inhibit rat erythroeyte hemolysis and lipid peroxidation of rat liver homogenate.CONCLUSION:The results strongly support the notion that EGb plays dual antioxidant roles: induction of antioxidant enzymes and direct reaction with ROS.

Objective To clone the gene of arginine kinase (AK) from Periplaneta americana, produce its recombinant protein and investigate its allergenicity. Method The cDNA of AK was cloned using specific primers from the total RNA of P. americana. The cloned gene was inserted into pMD18-T vector and digested by BamHI and HindⅢ. The cDNA was sequenced and subcloned into pET-28a expression vector. The cloned AK cDNA gene was expressed in Escherichia coli BL21 (DE3) by IPTG induction. The recombinant AK (rAK) was purified by metal (Ni2+) chelating affinity chromatography. Its allergenicity was examined by both Western blotting and enzyme-linked immunosorbent assay (ELISA). Result The cloned cDNA ORF sequence (Accession no. EU429466) contained 1 068 bp and encoded 365 amino acids. Its sequence homology with the published one (Accession no. AY563004) was 99.9% at nucleotide level. The allergen rAK was highly expressed in E. coli BL21 (DE3) as a soluble protein mainly with the molecular weight of about Mr 45 000 under induction of IPTG and purified by 6-His-tag purification system. Both in the non-denaturalization and denaturalization conditions, the recombinant allergen was identified as its affinity to IgE antibodies from the cockroach-allergic patient sera by Western blotting and ELISA. Conclusion The recombinant cockroach arginine kinase has been obtained with proper allergenicity.

Objective To investigate the knowledge level and needs in Xinjiang Uygur and Han patients with ulcerative colitis (UC), compare the differences between the two ethnic groups. Methods A total of 194 Uygur and Han UC patients were investigated with the general information questionnaire, the Crohn′s and Colitis Knowledge Score (CCKNOW) questionnaire, disease related knowledge needs questionnaire, to analyze the investigate results. Results CCKNOW score of Uygur and Han UC were (6.9±3.5) points and (9.2±3.1) points respectively, and Uygur was significantly lower than that of Han (Z=-2.831, P=0.005). Knowledge accuracy of dietary (30% and 63%) and drug (45% and 44%) were higher than general information (24% and 32%) and complications (12% and 30%) . Disease related knowledge needs score of Uygur and Han UC patients were (168.2±15.6) points and (155.4±17.2) points respectively, the score of Uygur patients was significantly higher than that of Han patients (t=4.429, P=0.001).The highest disease related knowledge needs score was (4.7±1.0) points of reproductive knowledge for Uygur UC patients, for Han patients was knowledge of daily life which was (4.8 ± 0.8) points. Conclusions The disease related knowledge level of Xinjiang Uygur and Han UC patients are relatively low, especially lack of general knowledge and diet knowledge, disease related knowledge of Uygur UC patients are lower than Han patients. The needs of the disease knowledge are very high for two ethnic groups, different one has different ethnic, gender, age, education level, living environment, hospitalization times, areas of disease related knowledge needs are also different. It is necessary to choose the targeted education content and suitable education way according to individual differences for nursing staff.

Objective To explore the association of anti-mullerian hormone(AMH)in seminal plasma and serum with sperm counts and energy for male.Methods For 215 cases of healthy male selected from our reproductive clinic,with women′s reason for infertility,seminal plasma and serum AMH were detected,as semen parameters(sperm density,living rate,vitality and malformation rate),6 items of serum sex hormone.In seminal plasma and serum AMH respectively as the dependent variable,using multiple line-ar regression model to explore its quantitative relation with semen parameters and sex hormone levels.Results 215 cases were en-rolled,aged 34.28±5.70 years,while the median of the seminal plasma AMH was 0.47,quartile 0.05-3.09 pmol/ejaculation.The median of the serum AMH was 53.07,quartile 32.32 -72.20 pmol/L.Through multiple linear regression analysis,after adjusted by age and BMI,the seminal plasma of AMH and total number of sperm,sperm concentration,dynamic motility,total sperm activi-ty,serum inhibin B were positively correlated(P <0.05);The correlation between sperm morphology and other serum sex hormone had no statistical significance(P >0.05);Serum AMH negatively correlated with serum FSH,with serum inhibin B positively(P <0.05);Seminal plasma in various parameters and other related serum sex hormone had no statistical significance(P >0.05).Conclu-sion The seminal plasma of AMH were positively correlated with sperm concentration,sperm counts,sperm vitality,with the asso-ciation for serum AMH not yet found.

Objective:To investigate effects of over-expression and suppression of HMGB1 on proliferation and invasion of endometrial carcinoma HEC-1A cell and underlying mechanisms.Methods:Over-expression or silence of HMGB 1 in HEC-1A cell lines were established by lentiviral vector containing HMGB 1 recombinant plasmid or by HMGB 1 shRNA,respectively.Cell counting kit-8,Transwell,and wound healing assay were used to analyze proliferation,invasion,and migration of HEC-1A cells,respectively.Western blot and reverse transcription-PCR (RT-PCR) were used to detect the expression of NF-κB,VEGF,and matrix metalloproteinase 2 (MMP2) in the cells.Results:Over-expression of HMGB1 promoted the proliferation,invasion,and migration of HEC1A cell,and up-regulated NF-κB,VEGF,and MMP2 expressions,while suppression of HMGB1 inhibited the proliferation,invasion,and migration of HEC-1A cells and down-regulated NF-κB,VEGF,and MMP2 expressions.Conclusion:HMGB1 plays an important role in proliferation,invasion,and migration of HEC1A cell via NF-κB/VEGF/MMP2 pathway.HMGB 1 might be a potential target for endometrial carcinoma therapy.

Objective To study the effects of folate(Fol) on proliferation,apoptosis and the antioxidative capacity of the NSCs under hypoxia in vitro.Method The NSCs taken from infant rats brain were cultured by serum-free medium in vitro and divided into four group which were normal control group(NC),hypoxia group(Hyp),Hpy+Fol-D(folate deficient)group and Hpy+Fol group.Except NC group,the other groups were cultured in hypoxia condition for 6 h.The proliferation of NSCs after hypoxia damage was detected by MTT test.The NSCs were collected after being cultured 6 d and then the activity of SOD and GSH-PX was examined.The expression of activated caspase-3 was detected by Western blot.Results Compared with NC group,the proliferation of NSCs and the activity of SOD and GSH-PX decreased significantly after hypoxia,while the expression of activated caspase-3 increased0 The proliferation of NSCs and the activity of SOD and GSH-PX in Hyp group were significantly higher than Hyp+Fol-D group but much lower than Hpy+Fol group(P

AIM To modify and improve a screening assay so that it becomes more convenient, economic and adaptable in China for high throughput screening of HIV fusion inhibitors targeting gp41. METHODS The original screening method reported by Jiang et al (J Virol. Methods 1999;80:85 96) was modified by: ① using a conformation specific monoclonal antibody to replace a polyclonal antibody for coating plates; ②simplifying the procedures; ③using parts of the reagents produced in China. RESULTS The modified screening assay is simpler, more convenient, and more economic than the original assay, but its sensitivity is comparable to and specificity is a little better than the original method. CONCLUSIONS The modified screening assay is more convenient and economic and can be used in China for high throughput screening of HIV fusion inhibitors from complex sample, such as phage display peptide libraries, microorganism fermentation liquids, herbs and other natural products.

BACKGROUND: There are currently many cryopreservation methods for the aminotic membrane, which have varying effects on the ultrastructure and biological activity of amniotic membrane, but on no one is effective.
OBJECTIVE: To compare the effects of different cryopreservation methods on the ultrastructure and viability of aminotic membrane and to seek the ideal cryopreservation method.
METHODS: Aminotic membrane separated from the fresh placenta was preserved respectively with deep-frozen cryopreservation and vitrification, and everyway was run for 3 and 6 months. Fresh aminotic membrane was used as control. The ultrastructure of aminotic membrane was observed by transmission electron microscopy, and the viability of aminotic membrane was assessed by microcomputer analysis system for biological oxygen consumption, and immunohistochemical staining combined with image analysis system was used for lactate dehydrogenase activity.
RESULTS AND CONCLUSION:After 3 and 6 months of crypreservation, the damage to the ultrastructure of aminotic membrane by vitreous cryopreservation was slighter than that of amniotic membrane cryopreserved at-80℃. Compared with the fresh aminotic membrane, the gray value of lactate dehydrogenase and partial pressure of oxygen were significantly decreased in the cryopreserved aminotic membrane by deep-frozen cryopreservation at 3 and 6 months (P < 0.05) and by vitreous cryopreservation at 6 months (P < 0.05), but there was no statisticaly significant difference in the change rate of oxygen partial pressure and the gray value of lactate dehydrogenase between the fresh aminotic membrane and the cryopreserved aminotic membrane by vitreous cryopreservation at 3 months. The present study led to the conclusion that vitreous cryopreservation protocol alows to not only maintain the integrity of AM, but also to preserve the viability of the cels. So the vitreous cryopreservation is superior to the deep-frozen cryopreservation for cryopreservation of aminotic membrane.

Objective To investigate the expression and clinical significance of DJ-1 protein in the endometrial carcinoma and delineate its functional roles in endometrial carcinoma progression.Methods Expression of DJ-1 was examined with immunohistochemical staining of 69 paraffin-embedded endometrial carcinoma samples who underwent surgery at Hunan province Tumor Hospital.Kaplan-Meier analysis was performed to describe the survival curve.Univariate and multivariate analyses were performed with COX proportional hazard model.Results (1) The expression of DJ-1 protein in endometrial carcinoma was significantly higher than that in normal endometrial tissues (P ＜ 0.05).(2) There was positive correlation between the high expression of DJ-1 protein in endometrial carcinoma samples and the pathological grade,the International Federation of Gynecology and Obstetrics (FIGO) stage,depth of myometrial invasion,or lymph node metastasis (P ＜ 0.05).(3) Kaplan-Meier analysis indicated that the cumulative 5-year survival rate in the low-DJ-1 expression group was 80.8％,whereas it was only 51.2％ in the high-DJ-1 expression group (P ＜ O.05).COX analysis indicated that the expression of DJ-1 was not the endometrial carcinoma independent prognostic factors for overall survival time (P ＞ 0.05).Conclusions The high expression of DJ-1 protein in endometrial carcinoma samples demonstrated a significantly positive correlation with the pathological grade,FIGO stage,depth of myometrial invasion,and lymph node metastasis.However,DJ-1 is not enough to be independent prognostic factor for overall survival time.

Objective To investigate the effects of different abstinent duration on brain functional network of heroin addicts at resting state.Methods Sixteen heroin addicts during protracted abstinence for 11-13 months (PA12 group) and twenty heroin addicts during protracted abstinence for 5-7 months (PA6 group) were recruited in the resting-state functional MRI study.Graph theoretical methods were applied to construct topological organization of whole brain network and nodes betweenness of the networks in all subjects,and the between-group differences were analyzed.The correlation of the node betweenness with the abstinence duration was conducted.Results There was no significant difference (P＞0.05) in the small world characteristic (γ≈1,λ》1) between two groups.Compared with the PA6 group,the PA12 group demonstrated significantly decreased nodal betweenness in regions of left parahippocampal gyrus,left precentral gyrus,and significantly enhanced nodal betweenness in regions of the left cuneus,left temporal and right middle occipital gyrus (all P＜0.05).Moreover,the betweenness of the left precentral gyrus (r =-0.52,P =0.001) and parahippocampal gyrus (r=-0.49,P=0.002) were negatively related with the abstinence duration,the betweenness of the right middle occipital was positively correlated with the abstinence duration (r=0.49,P=0.003).Conclusion The brain network small world topology of heroin addicts tend to be stable after 5-7 months of abstinence.Long-term abstinence may minimize the addicfs memory of drugs and potential drug seeking behavior,and recover addicts' visual spatial attention function.